What is the Ct standard deviation for the same sample in the whole pcr plate? - (Jun/01/2007 )

Hi.

Making a master mix with template and all - pipetting it equally in all the wells -

what is the observed Ct standard deviation and range?

I get on an older Opticom mashine 0.25 Ct deviation and all the samples are within +/- 0.4 Ct.

:-)

Peider

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I usually observe Ct deviances from 0.0 to 1.0 cycles from the same sample, depending on who did the pipetting, the template quality and the primer-combination used..... I usually accept Cts if they are within+/- 0.25 from each replicate....

Mike

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Hi all. I am a newbie here as well as to working in the lab. I am an undergrad but am working in a lab for the fall.


I am currently having issues of keeping my Ct values within tubes belonging to triplicates of the same primers/templates. It sounds as if it is a pipetting problem on my part, but I have taken great caution in performing equal mixing and whatnot.

How would the other factors posted by jadefalcon (template quality and primer-template combinations) contribute to a wider Ct range within a triplicate?

Thank you so much.

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